Effect and mechanism of Qingre Huashi decoction on drug-resistant Helicobacter pylori

Table 1 Composition of Qingre Huashi decoction

Authorities name	Scientific name	Plant part	Weight ratio (g)
Huanglian	Coptis chinensis Franch.	Rhizome	6
Zhizi	Gardenia jasminoides Ellis	Fruit	6
Pugongying	Taraxacum mongolicum Hand-Mazz.	Whole plant	15
Kuxingren	Prunus armeniaca L. var. ansu Maxim.	Seed	10
Doukouren	Amomun kravanh Pierre ex Gagnep.	Fruit	6
Yiyiren	Coix lacryma-jobi L. var. ma-yuen (Roman.) Stapf	Seed	15
Banxia	Pinellia ternata (Thunb) Breit.	Tuber	10
Cangzhu	Atractylodes lancea (Thunb.) DC.	Rhizome	10
Houpo	Magnolia Officinalis Rehd Et Wils.	Bark	10
Fuling	Poria Cocos (Schw.) Wolf.	Sclerotium	15
Cheqiancao	Plantago asiatica L.	Whole plant	15

Table 2 Information of 12 Helicobacter pylori-infected individuals

No.	Gender	Age	Gastroscopic diagnosis	Pathological diagnosis	13C
1	M	47	Superficial gastritis; duodenal bulb inflammation (mild)	Mild chronic gastritis in the gastric body, activity grade II, lymphoid follicle formation, and mucosal erosion	32.4
2	F	39	Superficial gastritis	Moderate chronic gastritis in the gastric antrum, activity grade II, and mucosal erosion	26.7
3	F	54	Chronic atrophic gastritis	Mild chronic gastritis in the gastric antrum, activity grade I, mild intestinal metaplasia. Mild chronic gastritis in the gastric fundus, and severe intestinal metaplasia	19.8
4	M	64	Reflux esophagitis (LA-A); superficial gastritis; duodenal bulb ulcer (H1 stage)	Moderate chronic gastritis in the gastric antrum, activity grade II, mucosal hyperplasia, lymphoid follicle formation, and fundic gland polyp in the gastric fundus	40.5
5	F	35	Chronic non-atrophic gastritis; bile reflux	Mild chronic inflammation in the gastric body, and activity grade II. Moderate chronic gastritis in the gastric antrum, activity grade II, and lymphoid follicle formation	42.2
6	F	42	Chronic non-atrophic gastritis; Helicobacter pylori infection	Moderate chronic inflammation in the gastric antrum, and activity grade II	27.5
7	M	32	Reflux esophagitis (LA-B); hiatal hernia; chronic atrophic gastritis C2	Moderate chronic gastritis in the gastric antrum, activity grade II	28.9
8	M	34	Chronic atrophic gastritis with intestinal metaplasia and gastric ulcer	Moderate chronic gastritis in the gastric antrum, activity grade II, mild intestinal metaplasia, mild atrophy, mucosal hyperplasia, lymphoid follicle formation. Mild chronic inflammation in the cardia, and activity grade I	24.3
9	F	35	Chronic superficial gastritis; bile reflux; duodenal bulb ulcer	Severe chronic inflammation in the gastric antrum, activity grade II, and lymphoid follicle formation	25.4
10	F	40	Hiatal hernia; chronic gastritis with intestinal metaplasia	Moderate chronic gastritis in the gastric antrum (site 1), activity grade II, mild intestinal metaplasia, and moderate glandular atrophy	17.3
11	F	46	Duodenal ulcer; chronic gastritis; bile reflux	NA	18.3
12	M	42	Superficial gastritis	Mild chronic gastritis in the gastric antrum, and focal lymphocyte aggregation	16.4

Table 3 Interpretation of biofilm production

Average optical density value	Biofilm production
≤ ODc	No
ODc < OD ≤ 2 × ODc	Weak
2 × ODc < OD ≤ 4 × ODc	Moderate
> 4 × ODc	Strong

Optical density cut-off value (ODc) = average optical density of negative control + 3 × SD of negative control. OD: Optical density; ODc: Optical density cut-off value.

Table 4 Primer sequences for quantitative polymerase chain reaction

Primer name	Sequence (5’→3’)	Primer name	Sequence (5’→3’)
16S rRNA-F	GCTAAGAGATCAGCCTATGTCC	Spot-F	TGTGCAACCTGTCGCTAATC
16S rRNA-R	TGGCAATCAGCGTCAGGTAATG	Spot-R	TTGGGCAAGATGTGGCTAAT
HP0605-F	AGCGCAAGAACTCAGTGTCA	LUXS-F	TGGATCACACYAAAGTCAAAG
HP0605-R	GCTTGGAGTTGTTGGGTGTT	LUXS-R	AAACCCCCACTTCAGACCA
HP1327-F	GCCAGGCTTGATGAAGAAAA	cagE-F	TTGAAAACTTCAAGGATAGGATAGAGC
HP1327-R	TTAGCCTGCTTGCCGTAAAT	cagE-R	GCCTAGCGTAATATCACCATTACCC
HP1489-F	TAGGCGCTCAAGTGGCTTAT	gluP-F	CCGCTGGTAATCCCTTTGTA
HP1489-R	TCAGATCGGGCAGATTTTTC	gluP-R	CTTGCATTATCGCCCATTTT
HP0971-F	TTACCGGCAAAGGGATACG	napA-F	TGAAGAGTTTGCGGACAT
HP0971-R	AAATTGGATCGCTCGTTGTATG	napA-R	AGAGTGGAAGCTCGTTTT

Table 5 Primers sequences for polymerase chain reaction-agarose gel electrophoresis

Primer name	Sequence (5’→3’)	Primer name	Sequence (5’→3’)
16S rRNA-F	GCTAAGAGATCAGCCTATGTCC	vacA m1/m2-F	CAATCTGTCCAATCAAGCGAG
16S rRNA-R	TGGCAATCAGCGTCAGGTAATG	vacA m1/m2-R	GCGTCAAAATAATTCCAAGG
CagA-F	GTTGATAACGCTGTCGCTTC	UreB-F	AGTAGCCCGGTGAACACAACATCCT
CagA-R	GGGTTGTATGATATTTTCCATAA	UreB-R	ATGCCTTTGTCATAAGCCGCTTGG
vacA s1/s2-F	ATGGAAATACAACAAACACAC	UreA-F	TGATGGGACCAACTCGTAACCGT
vacA s1/s2-R	CTGCTTGAATGCGCCAAAC	UreA-R	CGCAATGTCTAAGCGTTTGCCGAA

Table 6 Typing of Helicobacter pylori strains

No.	CagA	VacA	UreA	UreB	Helicobacter pylori strain type
1	+	+	+	-	I
2	+	+	+	+	I
3	+	+	+	+	I
4	+	+	+	+	I
5	+	+	+	-	I
6	-	+	+	+	I
7	+	+	+	+	I
8	-	+	+	+	I
9	+	+	+	+	I
10	+	+	+	+	I
11	+	+	+	+	I
12	-	+	+	+	I

+: Positive, -: Negative; Type I Helicobacter pylori strains: CagA and/or VacA (+); Type II Helicobacter pylori strains: CagA and VacA (-).

Table 7 Minimum inhibitory concentration of antibiotics against 12 clinical strains of Helicobacter pylori

Helicobacter pylori strain	AML (μg/mL)	CLR (μg/mL)	LEV (μg/mL)	MTZ (μg/mL)
1	0.016	16.0001	32.0001	64.001
2	0.064	24.0001	32.0001	256.001
3	0.016	8.0001	32.0001	256.001
4	0.064	16.0001	32.0001	256.001
5	0.064	32.0001	0.125	256.001
6	0.016	8.0001	3.0001	1.00
7	0.064	16.0001	32.0001	0.38
8	0.016	0.016	0.250	32.001
9	0.016	0.064	0.094	256.001
10	0.016	0.016	1.500	0.50
11	0.016	0.016	0.032	1.00
12	0.016	0.016	0.032	1.50

¹Antibiotic resistance.

MIC: Minimum inhibitory concentration; AML: Amoxicillin; CLR: Clarithromycin; LEV: Levofloxacin; MTZ: Metronidazole.

Table 8 Minimum inhibitory concentration of Qingre Huashi decoction against Helicobacter pylori

Helicobacter pylori strain	Concentration of QHD (mg/mL)
	NC (0)	16	32	64	128	256
1	+	+	+	-	-	-
2	+	+	+	-	-	-
3	+	+	+	-	-	-
4	+	+	+	-	-	-
5	+	+	+	-	-	-
6	+	+	+	-	-	-
7	+	+	+	-	-	-
8	+	+	-	-	-	-
9	+	+	+	-	-	-
10	+	+	+	-	-	-
11	+	+	+	-	-	-
12	+	+	+	-	-	-

MIC: Minimum inhibitory concentration; QHD: Qingre Huashi decoction; +: Existing colonies; −: No colony growth.