Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objective Exploring the efficacy and safety of botulinum toxin type A(BTX-A)combined with pulsed radiofrequency(PRF)in the treatment of postherpetic neuralgia(PHN).Methods A total of 80 patients with PHN were collected.They were randomly divided into experimental group(Group B)and control group(Group C),Group B was treated with BTX-A intradermal injection combined with PRF,and Group C was treated with lidocaine intradermal injection combined with PRF.Numeric pain score(NRS),Simplified McGill Pain Questionnaire(SF-MPQ)and Sleep Quality Score(QS)were used to assess the patients'pain level and sleep quality preoperatively,1,3,and 7 days postoperatively,and 1,2,and 3 months postoperatively.The patients'postoperative adverse reac-tions were collected.Interleukin-1β(IL-1β)and calcitonin gene-related peptide(CGRP)levels in patients'serum were measured preoperatively and 3 days postoperatively.Results The NRS scores,SF-MPQ scores,and QS scores of group B and group C were significantly lower at all postoperative time points compared to preoperative ones(P<0.05).The NRS and SF-MPQ score were significantly lower in group B at 1,2,and 3 months postoperatively compared with group C(P<0.05);and group B had significantly lower QS scores at 2 and 3 months postoperatively(P<0.05).The effective rate of pain relief at 3 months postoperatively in group B(90%)was statistically signifi-cant(P<0.05)compared with group C(56.7%).No serious adverse reactions occurred in either group.The levels of IL-1β and CGRP in serum of patients in both groups were significantly decreased at 3 days after surgery compared with the preoperative period,and the degree of decrease of IL-1β and CGRP in group B was more significant than that in group C(P<0.05).Conclusion BTX-A combined with PRF treatment for PHN can effectively reduce its pain level,improve the quality of sleep,and is safe.

Abstract: Objective　To investigate the molecular characteristics and drug resistance of non-O1/non-O139 Vibrio cholerae in Zhongshan City, and to provide laboratory basis for cholera prevention and control. Methods　The strains of non-O1/non-O139 Vibrio cholerae isolated from sporadic patients and aquatic products from 2015 to 2021 in Zhongshan city were collected. The identification and cluster analysis of the strains were analyzed by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), the ctxA virulence gene of strains were detected by real-time fluorescence quantitative PCR, the cluster analysis of the strains was analyzed by pulsed-field gel electrophoresis (PFGE), and the drug resistance of the strains were analyzed by microbroth dilution method. Results　From 2015 to 2021, 33 strains of non-O1/non-O139 Vibrio cholerae were isolated from Zhongshan City, including 28 strains from sporadic patients and 5 strains from aquatic products. Through MALDI-TOF-MS identification, 33 strains of non-O1/non-O139 Vibrio cholera can be identified to the level of species, and the identification results were all Vibrio cholerae. Among 33 non-O1/non-O139 Vibrio cholerae strains, 1 strain carried the ctxA virulence gene. The drug-resistant strains accounted for 69.7% (23/33), and the multidrug resistant strains accounted for 18.2% (6/33). A total of 7 kinds of drug resistance spectrum were produced, including 3 kinds of multidrug resistant spectrum, and showed drug resistance to 8 antibiotics, among which the resistance rates to streptomycin, cefazolin and compound sulfamethoxazole were above 30%. The 33 strains of non-O1/non-O139 Vibrio cholerae were divided into 32 PFGE fingerprints with a similarity ranging from 61.7% to 100%. MALDI-TOF-MS cluster analysis divided 33 non-O1/non-O139 Vibrio cholerae strains into two clusters. Conclusions The results of molecular typing of non-O1/non-O139 Vibrio cholerae in Zhongshan City presented diversity, and no significant correlation was found between PFGE and MALDI-TOF-MS cluster analysis. The strains demonstrated various degrees of resistance to certain antibiotics, and there were multidrug-resistant and toxigenic strains. Therefore, it is necessary to alert to the harmfulness of non-O1/non-O139 Vibrio cholerae and enhance monitoring.

【Objective 】 To explore the influencing factors of Yang deficiency constitution in traditional Chinese medicine (TCM) from the perspective of mathematics with the use of calculation formulas, so as to protect patients from getting diseases caused by Yang deficiency constitution and provide suggestions for TCM disease prevention. 【Methods】  Based on the classification and determination criteria of TCM constitution implemented by China Association of Chinese Medicine, data for 24 solar terms from May 5, 2020(Start of Summer) to April 20, 2021 (Grain Rain) for the identification of Yang deficiency were collected by mobile constitution identification system. The grey correlation analysis method was used to determine the grey correlation degree of the factors influencing Yang deficiency constitution. In addition, a random forest model was constructed for the verification of the results from the grey correlation analysis, and for the evaluation of correlation degree between Yang deficiency constitution and its influencing factors. 【Results】  A total of 16 259 sets of data were collected from healthy or sub-healthy individuals aged from 18 to 60 years living in the central and northeastern parts of Sichuan Province(China) for the identification of TCM constitutions. After screening and preprocessing, a total of 544 sets of data for the identification of Yang deficiency constitution, involving 18 aspects of factors influencing Yang deficiency constitution. The results of the grey correlation analysis showed that there were 12 influencing factors whose grey correlation degree with Yang deficiency constitution was greater than 0.6. The accuracy of these 12 influencing factors with the training set and validation set of the Yang deficiency constitution random forest model were 98.39% and 93.12%, respectively. 【Conclusion】  In the sample data selected in this paper, grey correlation analysis is the appropriate technology to analyze the influencing factors of Yang deficiency constitution. It provides a new idea and a new methodological reference for the research and analysis of the influencing factors of TCM constitution.

ObjectiveTo discuss the diagnostic methods of global developmental delay caused by 10q24.3 heterozygous loss. MethodsA retrospective analysis was conducted on the clinical data of one child with global developmental delay， and the results of low depth whole-genome copy number variation sequencing （CNVseq） and family whole exome sequencing （WES） of the child and his parents. ResultsThe patient was a 10-month-old male with developmental retardation in four areas， with some special features （ocular hypertelorism， strabismus， flat nose bridge， protruding forehead， cleft palate， high palatal arch， etc.） and hypotonia of limbs. The CNVseq and WES test showed that the patient had new 10q24.3 heterozygosis loss. Because this region contains the gene SUFU associated with basal cell nevus syndrome and the gene CNNM2 associated with hypomagnesemia， seizures， and mental retardation， and the gene TRIM8 associated of Focal segmental glomerulosclerosis with neurodevelopmental syndrome， we speculated that the cause of the disease in the child was highly related to the heterozygosity deletion of SUFU gene and CNNM2 gene and TRIM8 gene. ConclusionGenetic testing should be improved as soon as possible for children with global developmental delay and special facial manifestations， so as to make clear diagnosis and to judge prognosis.

ObjectiveA previous multicenter， single-arm phase Ⅱ clinical study evaluated the efficacy and safety of the PD-1 inhibitor camrelizumab in combination with the antiangiogenic drug apatinib in patients with recurrent/metastatic cervical cancer. In the current study， we aimed to report the patient-reported outcomes （PRO） in patients with advanced cervical cancer. MethodsThe researchers filled out quality of life questionnaire FACT-Cx with the patients face-to-face at the baseline and every two treatment cycles. The patients provided the questionnaire until disease progression. For patients who continued the study， the data cutoff date for the questionnaire was Oct12， 2020. ResultsIn this study， 41 patients who received treatment finished at least one FACT-Cx questionnaire. The scores of FACT-Cx questionnaire at baseline， the first 6 cycles， and after 6 cycles of treatment （range， 7-15cycle） were 114.53±18.23， 124.08±15.23 and 132.82±21.97， respectively. The scores of questionnaire at the first 6 cycle and after 6 cycles of treatment were significantly increased when compared with those at baseline （P＜0.05）. With respect to each dimension of the FACT-Cx questionnaire， physical status （such as mental state， limb pain）， emotional status （such as sadness， tension）， functional status （such as daily activities） and additional concerns （such as sexual function）， the average scores of these dimensions was significantly higher after 6 treatment cycles than those at baseline （P<0.05）. Social/family status dimensions （such as family relationships） did not change significantly. In the first 6 cycles， the incidence of grade 3 to 4 adverse events （AEs） was 71.11%. However， the incidence of grade 3 to 4 AEs decreased significantly to 25.0% after 6 treatment cycles， with the frequencies of hypertension and fatigue decreasing significantly. ConclusionsCamrelizumab combined with apatinib significantly improves the overall quality of life of patients with recurrent/metastatic cervical cancer. The scores of FACT-Cx questionnaire in multiple dimensions， such as physiological status， emotional status， functional status， and the additional status were significantly increased. Moreover， the questionnaire scores still increase after 6 treatment cycles compared with those of the first 6 cycles， indicating that the improvement of quality of life is durable.